Abstract
Aim. Alfalfa is a rich source of phytoestrogens, among them coumestrol which shows strong estrogenic activity that can adversely affect the health of domestic animals. The aim of the study was to determine the variation in coumestrol content in leaves of alfalfa breeding populations, present in the breeding program of Agricultural Institute Osijek in Croatia. Method. Twelve alfalfa populations were selected based on their high forage yield and good persistence. Coumestrol was extracted using acidifi ed methanol as an organic solvent from lyophilized and ground alfalfa leaves, while for detection and quantifi cation was used. Results. Signifi cant differences were observed between the studied populations with average coumestrol content of 435.67 mg/kg of dry matter (DM). The highest content of coumestrol was determined in breeding population Rs-21 (619.53 mg/kg of DM). Conclusions. Populations Rs-33 and Rs-20 had the lowest coumestrol content (82.18 and 86.58 mg/kg, respectively) and present a potential breeding source for creat- ing new contemporary cultivars with decreased coumestrol content
Highlights
Alfalfa (Medicago sativa L.) is the most significant perennial forage legume in the production of high quality and nutritionally valuable fodder for livestock feed, primarily ruminants (Vasileva, 2013; Tucak et al, 2018)
The samples were frozen at −80 °C, lyophilized, ground to a powder from which coumestrol was extracted and quantified by Reverse phase (RP)-HPLC analysis, using a Perkin Elmer LC 200 HPLC system liquid chromatograph, with DAD detector and RP-C18 column, according to the methodology previously described in detail in Tucak et al (2020)
The lowest coumestrol content was obtained from the breeding population Rs-33 (82.18 mg/kg of dry matter (DM)), which was not significantly lower than the value determined for the Rs-20 population
Summary
Alfalfa (Medicago sativa L.) is the most significant perennial forage legume in the production of high quality and nutritionally valuable fodder for livestock feed, primarily ruminants (Vasileva, 2013; Tucak et al, 2018). Despite its exceptional value in livestock systems, alfalfa, like many other legume species and varieties, may contain phytoestrogens, the most interesting of which is coumestrol, which express strong estrogenic activity that can adversely affect the reproductive health of domestic animals (Hloucalova et al, 2016; Reed, 2016). In Medicago species, coumestrol production is less influenced by genetic factors, and plant selection/ breeding usually will not be effective for controlling its content (Reed, 2016). Wide variation in coumestrol content is affected by numerous factors such as environmental conditions (temperature, rainfall, humidity, sunlight), cultivar, stressed conditions (especially by fungal diseases, insect infestation and nutrient deficiencies), stages of maturity, plant parts analyzed, management strategies and stand age of crops (Seguin et al, 2004; Aragadvay-Yungán et al, 2017; Fields et al, 2018b; Fields et al, 2019; Tucak et al, 2020). Reduced coumestrol accumulation could be achieved by creating and selecting new modern yielding and fungal resistant alfalfa cultivars
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